Generating Kidney from Stem Cells.

Human kidney tissue can now be generated via the directed differentiation of human pluripotent stem cells. This advance is anticipated to facilitate the modeling of human kidney diseases, provide platforms for nephrotoxicity screening, enable cellular therapy, and potentially generate tissue for renal replacement. All such applications will rely upon the accuracy and reliability of the model and the capacity for stem cell-derived kidney tissue to recapitulate both normal and diseased states. In this review, we discuss the models available, how well they recapitulate the human kidney, and how far we are from application of these cells for use in cellular therapies.

Sero-molecular epidemiology of Japanese encephalitis virus in swine population of western Uttar Pradesh, India: Unraveling the geographical expansion of the virus.

BACKGROUND & OBJECTIVES: Swine is a good sentinel for forecast of Japanese encephalitis virus (JEV) outbreaks in humans. The present study was envisaged with objectives to know the sero-conversion period of JEV and to assess the prevalence of JEV in swine population of western Uttar Pradesh state of India. METHODS: A total of 252 swine serum samples were screened using IgM ELISA over the period of one year to determine the sero-conversion rate and compared seasonally to check the transmission peak of virus. Further, 321 swine blood and serum samples were collected from all seven divisions of western Uttar Pradesh to determine prevalence of JEV using real time RT-PCR and ELISA. RESULTS: Seasonal sero-conversion rate was high during monsoon and post-monsoon (32%) followed by winter (22.91%) and summer (10.71%) seasons. The sero-conversion was observed in all months indicating viral activity throughout the year in the region. The low degree of correlation was found between meteorological variables (day temperature, rainfall) and sero-conversion rate. A total of 52 samples (16.19%) were found positive by real time RT-PCR while sero-positivity of 29.91% was observed using IgG and IgM ELISA(s). The overall prevalence of JEV was 39.25%. INTERPRETATION & CONCLUSION: The presence of JEV was recorded throughout the year with peak occurrence during monsoon and post-monsoon season indicating that virus has spread its realm to western region of the state. The information generated in the present study will aid in initiating timely vector control measures and human vaccination program to mitigate risk of JEV infection in the region.

Kidney micro-organoids in suspension culture as a scalable source of human pluripotent stem cell-derived kidney cells.

Kidney organoids have potential uses in disease modelling, drug screening and regenerative medicine. However, novel cost-effective techniques are needed to enable scaled-up production of kidney cell types in vitro We describe here a modified suspension culture method for the generation of kidney micro-organoids from human pluripotent stem cells. Optimisation of differentiation conditions allowed the formation of micro-organoids, each containing six to ten nephrons that were surrounded by endothelial and stromal populations. Single cell transcriptional profiling confirmed the presence and transcriptional equivalence of all anticipated renal cell types consistent with a previous organoid culture method. This suspension culture micro-organoid methodology resulted in a three- to fourfold increase in final cell yield compared with static culture, thereby representing an economical approach to the production of kidney cells for various biological applications.

Dental Management Considerations for Patients with Cardiovascular Disease-A Narrative Review.

Dental therapists encounter patients with various systemic diseases of which cardiovascular disease (CVD) patients form a significant segment. Relation between oral health and cardiac diseases has been well established. Common cardiac disorders encountered in a dental practice include arterial hypertension, heart failure, ischemic heart disease, cardiac arrhythmias, infective endocarditis, stroke, and cardiac pacemaker. Patients with CVDs pose a significant challenge to dental therapy. These patients need special considerations and an adequate understanding of the underlying cardiovascular condition to provide safe and effective dental treatment. Based on the cardiac condition, an appropriate modification in dental care is crucial. A multidisciplinary approach including the patient's cardiologist can potentially reduce complications and improve dental treatment results. This review aims at unfolding the risks associated with the dental management of a cardiac patient and outlines the measures to be undertaken for optimum dental treatment.

Recapitulating kidney development: Progress and challenges.

Decades of research into the molecular and cellular regulation of kidney morphogenesis in rodent models, particularly the mouse, has provided both an atlas of the mammalian kidney and a roadmap for recreating kidney cell types with potential applications for the treatment of kidney disease. With advances in both our capacity to maintain nephron progenitors in culture, reprogram to kidney cell types and direct the differentiation of human pluripotent stem cells to kidney endpoints, renal regeneration via cellular therapy or tissue engineering may be possible. Human kidney models also have potential for disease modelling and drug screening. Such applications will rely upon the accuracy of the model at the cellular level and the capacity for stem-cell derived kidney tissue to recapitulate both normal and diseased kidney tissue. In this review, we will discuss the available cell sources, how well they model the human kidney and how far we are from application either as models or for tissue engineering.

Original study: early patient-reported functional outcome of all-inside ACL reconstruction as compared to anteromedial portal technique.

BACKGROUND: Anteromedial portal technique (AMP) using hamstring autograft is a popular technique of arthroscopic ACL reconstruction allowing anatomical placement of femoral. In this technique, a cortical suspensory fixation of graft is used on the femoral side and interference screw fixation on the tibial side using a complete bone tunnel. All-inside translateral technique is a recently introduced technique which uses a closed loop of quadrupled semitendinosus graft with an adjustable cortical suspensory fixation on both sides allowing optimum tensioning of graft and near-complete filling of retrosockets created by the flip cutter on both femoral and tibial sides. With its proposed but unproven benefits, our study was planned to compare the two techniques. METHODS: A total of 80 young active males requiring ACL reconstruction surgery were equally randomized to AMP and All-inside technique. The primary objective of the study was to compare the ability to return to pre-injury level of activity using Tegner activity scale and patient-reported outcome using new Knee Society Score (KSS) at two years of follow-up. RESULTS: The mean improvement in Tegner score was significantly better (p = 0.0005) in all-inside group (2.34 ± 0.97) as compared to AMP group (1.5 ± 1.30). Among components of new KSS, patient satisfaction was better in all-inside group. CONCLUSION: All-inside ACL reconstruction provides a better chance of return to pre-injury level of activity with accompanied patient satisfaction as compared to AMP technique at two years of follow-up. LEVEL OF EVIDENCE: Level I, therapeutic study.

A toxicogenomic approach for the risk assessment of the food contaminant acetamide.

Acetamide (CAS 60-35-5) is detected in common foods. Chronic rodent bioassays led to its classification as a group 2B possible human carcinogen due to the induction of liver tumors in rats. We used a toxicogenomics approach in Wistar rats gavaged daily for 7 or 28 days at doses of 300 to 1500 mg/kg/day (mkd) to determine a point of departure (POD) and investigate its mode of action (MoA). Ki67 labeling was increased at doses ≥750 mkd up to 3.3-fold representing the most sensitive apical endpoint. Differential gene expression analysis by RNA-Seq identified 1110 and 1814 differentially expressed genes in male and female rats, respectively, following 28 days of treatment. Down-regulated genes were associated with lipid metabolism while up-regulated genes included cell signaling, immune response, and cell cycle functions. Benchmark dose (BMD) modeling of the Ki67 labeling index determined the BMD10 lower confidence limit (BMDL10) as 190 mkd. Transcriptional BMD modeling revealed excellent concordance between transcriptional POD and apical endpoints. Collectively, these results indicate that acetamide is most likely acting through a mitogenic MoA, though specific key initiating molecular events could not be elucidated. A POD value of 190 mkd determined for cell proliferation is suggested for risk assessment purposes.

The gut flora modulates intestinal barrier integrity but not progression of chronic kidney disease in hyperoxaluria-related nephrocalcinosis.

BACKGROUND: Dysbiosis, bacterial translocation and systemic inflammation have been found to be associated with human and experimental forms of chronic kidney disease (CKD), but the functional contribution of the intestinal microbiota to CKD-related intestinal barrier dysfunction and CKD progression is unknown, especially in CKD secondary to hyperoxaluria and nephrocalcinosis. METHODS: C57BL/6N mice fed an oxalate-rich diet for either 10 or 20 days developed reversible or progressive kidney disease, respectively. RESULTS: Oxalate-induced CKD manifested as azotaemia, renal anaemia and hyperkalaemia. CKD was associated with persistent dysbiosis and intestinal barrier dysfunction. Local as well as systemic inflammation was evident and partially persisted despite better renal function after returning to an oxalate-free diet, indicating some innate immune memory. Eradication of the microbiota with a combination of antibiotics improved intestinal barrier function but had no effect on renal function, nephrocalcinosis, kidney remodelling and atrophy compared with control mice not receiving antibiotics. CONCLUSIONS: Together, in chronic oxalate nephropathy, the intestinal microbiota contributes to the CKD-related dysfunction of the intestinal barrier but not to the progression of nephrocalcinosis itself, as well to its related kidney atrophy and excretory dysfunction.

HIV Associated Risk Factors for Ischemic Stroke and Future Perspectives.

Although retroviral therapy (ART) has changed the HIV infection from a fatal event to a chronic disease, treated HIV patients demonstrate high prevalence of HIV associated comorbidities including cardio/cerebrovascular diseases. The incidence of stroke in HIV infected subjects is three times higher than that of uninfected controls. Several clinical and postmortem studies have documented the higher incidence of ischemic stroke in HIV infected patients. The etiology of stroke in HIV infected patients remains unknown; however, several factors such as coagulopathies, opportunistic infections, vascular abnormalities, atherosclerosis and diabetes can contribute to the pathogenesis of stroke. In addition, chronic administration of ART contributes to the increased risk of stroke in HIV infected patients. Concurrently, experimental studies in murine model of ischemic stroke demonstrated that HIV infection worsens stroke outcome, increases blood brain barrier permeability and increases neuroinflammation. Additionally, residual HIV viral proteins, such as Trans-Activator of Transcription, glycoprotein 120 and Negative regulatory factor, contribute to the pathogenesis. This review presents comprehensive information detailing the risk factors contributing to ischemic stroke in HIV infected patients. It also outlines experimental evidence demonstrating the impact of HIV infection on stroke outcomes, in addition to possible novel therapeutic approaches to improve these outcomes.

New synthetic coumarinolignans as attenuators of pro-inflammatory cytokines in LPS-induced sepsis and carrageenan-induced paw oedema models.

OBJECTIVES: The aim of the study was to explore the inhibition efficacy of new synthetic coumarinolignans (SCLs) against the secretion of pro-inflammatory cytokines in two in vivo models of inflammation. METHODS: Four SCLs 1-4 were screened for their pro-inflammatory cytokine inhibitory potential through oral administration at a dose of 50 mg/kg body weight in lipopolysaccharide-induced mouse endotoxaemia and carrageenan-induced mouse paw oedema models. Levels of pro-inflammatory cytokines (IL-1ß, TNFα and IL-6) in blood and paw tissue samples were estimated using ELISA. Paw oedema was measured using a plethysmometer. Results were compared with a natural coumarinolignan, cleomiscosin A (5), and the structure-activity relationship (SAR) was interpreted. RESULTS AND DISCUSSION: Compound 2 had the greatest potential in the endotoxaemia model, exhibiting 66.41%, 62.56% and 43.15% inhibition of plasma IL-1ß, TNFα and IL-6 secretions, respectively. Further dose-dependent study revealed its anti-inflammatory potential even at dose of 10 mg/kg body weight with 24.42% decline in the level of IL-1ß. Nevertheless, SCLs 1, 3 and 4 showed marked inhibitory activity with 57.54%, 51.48% and 62.46% reduction in the levels of IL-1ß, respectively. Moreover, compound 2 decreased the plasma TNFα and IL-1ß levels to 50.03% and 36.58% along with the reduction of paw oedema volume in the local inflammation induced by carrageenan. All compounds including cleomiscosin A (5) were more effective against IL-1ß. By studying SAR, the presence of dihydroxyl groups in the phenyl ring of lignans was identified to be essential for the activity. Also, esterification of lignans and presence of a 4-methyl substituent in the coumarin nucleus were found to play some role in enhancing the activity. CONCLUSION: All four SCLs, especially compound 2, have shown vast potential to emerge out as promising anti-inflammatory drugs.

Diode laser-assisted management of intraoral soft tissue overgrowth: a case series.

The conventional method of managing intraoral soft tissue overgrowth involves surgical excision using a scalpel under local anesthesia. These procedures are often associated with excessive bleeding that leads to complications. Now, minimally invasive methods such as laser excision are available for the management of intraoral soft tissue lesions. The objective of the present case series is to describe the use of a 980-nm diode laser to treat 4 cases of soft tissue enlargement of the oral cavity. Two women and 2 men with an age range of 21-73 years presented with chief complaints of intraoral swelling. The overgrowth was located on the tongue in 2 patients and in the gingival region in 2 patients. In all 4 patients, the overgrown tissues were excised with a 980-nm gallium-arsenide diode laser at a power of 1.0-4.0 W in continuous-wave contact mode. Excised tissues were sent for histopathologic examination to confirm the clinical diagnosis. Each lesion was excised with adequate hemostasis in 15-20 minutes. At the recall examination after 10 days, patients did not report any postoperative complications. There was no sign of recurrence in any patient at the 3-month follow-up. Based on the histopathologic reports, 2 lesions were diagnosed as pyogenic granuloma, 1 as capillary hemangioma (a form of pyogenic granuloma), and 1 as peripheral ossifying fibroma. The results of this limited case series suggest that a 980-nm diode laser may be an advantageous surgical modality for treating benign proliferative lesions of the oral mucosa with negligible postoperative adverse sequelae.

Immunomodulatory Molecule IRAK-M Balances Macrophage Polarization and Determines Macrophage Responses during Renal Fibrosis.

Activation of various innate immune receptors results in IL-1 receptor-associated kinase (IRAK)-1/IRAK-4-mediated signaling and secretion of proinflammatory cytokines such as IL-12, IL-6, or TNF-α, all of which are implicated in tissue injury and elevated during tissue remodeling processes. IRAK-M, also known as IRAK-3, is an inhibitor of proinflammatory cytokine and chemokine expression in intrarenal macrophages. Innate immune activation contributes to both acute kidney injury and tissue remodeling that is associated with chronic kidney disease (CKD). Our study assessed the contribution of macrophages in CKD and the role of IRAK-M in modulating disease progression. To evaluate the effect of IRAK-M in chronic renal injury in vivo, a mouse model of unilateral ureteral obstruction (UUO) was employed. The expression of IRAK-M increased within 2 d after UUO in obstructed compared with unobstructed kidneys. Mice deficient in IRAK-M were protected from fibrosis and displayed a diminished number of alternatively activated macrophages. Compared to wild-type mice, IRAK-M-deficient mice showed reduced tubular injury, leukocyte infiltration, and inflammation following renal injury as determined by light microscopy, immunohistochemistry, and intrarenal mRNA expression of proinflammatory and profibrotic mediators. Taken together, these results strongly support a role for IRAK-M in renal injury and identify IRAK-M as a possible modulator in driving an alternatively activated profibrotic macrophage phenotype in UUO-induced CKD.

Influence of polycystic ovary syndrome on the periodontal health of Indian women visiting a secondary health care centre.

OBJECTIVES: Periodontal disease and polycystic ovary syndrome (PCOS) share risk factors like obesity, insulin resistance, and dyslipidemia, along with evidence of chronic inflammation in the two conditions. Evaluating the influence of PCOS on periodontal health would, therefore, identify a possible association. MATERIALS AND METHODS: Sixty women, divided into equal groups of PCOS and healthy patients, were clinically examined for periodontal parameters like probing depth (PD), plaque index (PI), modified gingival index (mGI), and bleeding on probing (BOP). Fasting blood sugar (FBS), insulin (FI), triglycerides (TG), and free testosterone along with serum and gingival crevicular fluid (GCF) levels of malondialdehyde (MDA) and myeloperoxidase (MPO) were the biochemical parameters evaluated. RESULTS: Women with PCOS had statistically significant differences in mGI, PI, testosterone, FBS, and TG when compared with healthy women (p < 0.05). MDA levels in serum and GCF between women with PCOS and controls were also significantly different. BOP and mGI showed a moderate positive correlation (r = 0.45 and 0.44) with serum levels of MDA. Relatively greater gingival inflammation was observed in patients with PCOS compared to healthy controls, independent of the risk factors present. CONCLUSION: PCOS seemed to have an impact on gingival inflammation, in addition to the effect of dental plaque and other local factors in the oral cavity, in PCOS patients when compared with healthy individuals. CLINICAL RELEVANCE: Women diagnosed with PCOS may have probabaility of co-existing gingival inflammation. Therefore, emphasis on medical treatment for PCOS and periodic screening for periodontal disease may be warranted.

Assessment of Pyrexia and Associated Sickness Behavior in Patients with Chronic Periodontitis.

BACKGROUND AND OBJECTIVES: The purpose of this study was to assess pyrexia and sickness behavior such as anxiety, depression, lethargy, and weight loss in subjects with chronic periodontitis, and evaluate inflammatory mediators such as erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) in the patients with fever. METHODS: This was a cross-sectional study that consisted of 150 chronic periodontitis and 150 healthy subjects. Sublingual and subgingival temperatures were assessed using a digital temperature probe. Associated sickness behavior was assessed for all the subjects. Pathological tests, i.e., ESR and CRP were done for subjects with fever. RESULTS: Evaluating the presence of fever with the severity of periodontal disease, 66.7% of the subjects with fever were in the group diagnosed with severe periodontitis, 20.4% had with moderate periodontitis, and 5.3% were in the healthy group. Subjects diagnosed with periodontitis comprised 66.2% of the subjects with higher (i.e., > 36.3°C) subgingival temperatures and healthy subjects made up the remaining 33.8% (p < 0.001). A correlation between the sublingual and subgingival temperature with the Pearson ρ correlation coefficient of 0.227 (p < 0.001) was observed. A statistically significant mean value of 37.05 ± 11.24 of ESR and 1.59 ± 1.11 mg/L of CRP was reported amongst the subjects with fever (p < 0.001). The association of sickness behavior with the severity of periodontitis was found to be significant: depression 40.006, anxiety 50.857, reported weight loss 76.463, and lethargy 141.581 (p < 0.001). DISCUSSION AND CONCLUSION: The study demonstrated that there is a significant increase in the sublingual temperature amongst patients with severe chronic periodontitis. The subgingival temperature has a positive correlation with the sublingual temperature. There was a linear trend of an association of sickness behavior with the severity of chronic periodontitis. A significant increase in the circulating inflammatory mediators, CRP and ESR, were noticed in subjects with elevated body temperature.

Impact of oral potentially malignant disorders on quality of life.

BACKGROUND: Oral potentially malignant disorders (OPMDs) could have a significant psychological impact on patients, principally because of the unknown risk of malignant transformation, while the physical and functional impairments could differ. This study aimed to assess the impact of three different OPMDs and their disease stages on the quality of life (QoL) of affected patients. METHODS: Oral leukoplakia (OL), oral lichen planus (OLP) and oral submucous fibrosis (OSF) patients who were undergoing treatment at an oral medicine clinic of a dental teaching hospital in India were the study population. All subjects completed the recently developed OPMDQoL questionnaire and a short form 12 item (version 2) health survey questionnaire (SF-12v2). OPMDQoL questionnaire consists of 20 items over four dimensions. A higher score denotes poor OHRQoL. SF-12v2 has two components, a Physical Component Summary (PCS) and Mental Component Summary (MCS). RESULTS: A total of 150 subjects (50 each of OL, OLP and OSF) participated. OL patients (37.7 ± 7.9) reported significantly better OPMDQoL scores than OLP (47.3 ± 5.8) and OSF (45.4 ± 9.2) patients. OLP patients reported significant problems in obtaining a clear diagnosis for their condition, more so than the other OPMDs. OL patients reported fewer problems for the dimension, "physical impairment and functional limitations" than the OLP and OSF patients. A significant trend was observed with the overall OPMDQoL and MCS, deteriorating as the disease stage increased. CONCLUSIONS: OLP and OSF have a significant impact on the QoL of affected individuals: OL less so. Increasing stage of the disease is associated with worsening QoL.

Histones and Neutrophil Extracellular Traps Enhance Tubular Necrosis and Remote Organ Injury in Ischemic AKI.

Severe AKI is often associated with multiorgan dysfunction, but the mechanisms of this remote tissue injury are unknown. We hypothesized that renal necroinflammation releases cytotoxic molecules that may cause remote organ damage. In hypoxia-induced tubular epithelial cell necrosis in vitro, histone secretion from ischemic tubular cells primed neutrophils to form neutrophil extracellular traps. These traps induced tubular epithelial cell death and stimulated neutrophil extracellular trap formation in fresh neutrophils. In vivo, ischemia-reperfusion injury in the mouse kidney induced tubular necrosis, which preceded the expansion of localized and circulating neutrophil extracellular traps and the increased expression of inflammatory and injury-related genes. Pretreatment with inhibitors of neutrophil extracellular trap formation reduced kidney injury. Dual inhibition of neutrophil trap formation and tubular cell necrosis had an additive protective effect. Moreover, pretreatment with antihistone IgG suppressed ischemia-induced neutrophil extracellular trap formation and renal injury. Renal ischemic injury also increased the levels of circulating histones, and we detected neutrophil infiltration and TUNEL-positive cells in the lungs, liver, brain, and heart along with neutrophil extracellular trap accumulation in the lungs. Inhibition of neutrophil extracellular trap formation or of circulating histones reduced these effects as well. These data suggest that tubular necrosis and neutrophil extracellular trap formation accelerate kidney damage and remote organ dysfunction through cytokine and histone release and identify novel molecular targets to limit renal necroinflammation and multiorgan failure.

Hyperoxaluria Requires TNF Receptors to Initiate Crystal Adhesion and Kidney Stone Disease.

Intrarenal crystals trigger inflammation and renal cell necroptosis, processes that involve TNF receptor (TNFR) signaling. Here, we tested the hypothesis that TNFRs also have a direct role in tubular crystal deposition and progression of hyperoxaluria-related CKD. Immunohistochemical analysis revealed upregulated tubular expression of TNFR1 and TNFR2 in human and murine kidneys with calcium oxalate (CaOx) nephrocalcinosis-related CKD compared with controls. Western blot and mRNA expression analyses in mice yielded consistent data. When fed an oxalate-rich diet, wild-type mice developed progressive CKD, whereas Tnfr1-, Tnfr2-, and Tnfr1/2-deficient mice did not. Despite identical levels of hyperoxaluria, Tnfr1-, Tnfr2-, and Tnfr1/2-deficient mice also lacked the intrarenal CaOx deposition and tubular damage observed in wild-type mice. Inhibition of TNFR signaling prevented the induced expression of the crystal adhesion molecules, CD44 and annexin II, in tubular epithelial cells in vitro and in vivo, and treatment with the small molecule TNFR inhibitor R-7050 partially protected hyperoxaluric mice from nephrocalcinosis and CKD. We conclude that TNFR signaling is essential for CaOx crystal adhesion to the luminal membrane of renal tubules as a fundamental initiating mechanism of oxalate nephropathy. Furthermore, therapeutic blockade of TNFR might delay progressive forms of nephrocalcinosis in oxalate nephropathy, such as primary hyperoxaluria.

Ninety-Day Morbidity and Mortality in Risk-Screened and Optimized Patients Undergoing Two-Team Fast-Track Simultaneous Bilateral TKA Compared With Unilateral TKA-A Prospective Study.

BACKGROUND: Simultaneous bilateral total knee arthroplasty (SBTKA) offers significant socioeconomic benefits. However, retrospective studies and public health data show increased mortality and morbidity rates in patients undergoing SBTKA compared with those undergoing unilateral TKA (UTKA), and there have been recommendations against the use of SBTKA. High-volume centers, which feature careful patient selection and fast-tracked surgery, continue to perform SBTKA and have published their results in favor of the procedure. However, the quality of evidence remains poor. METHODS: We prospectively examined 90-day morbidity and mortality of SBTKA compared with UTKA in risk-screened and optimized patients in our high-volume joint replacement facility. A total of 1200 consecutive patients were recruited in each arm. RESULTS: Ninety-day mortality was higher in SBTKA patients than in UTKA patients (0.58% vs 0.42%, respectively; P = .5646). Overall procedure-related complications were significantly higher in the SBTKA group (7.25% vs 4.42%, respectively; P = .0034). The relative risk of cardiovascular complications in SBTKA patients was 6.5 times higher than that in UTKA patients (1.08% vs 0.17%, respectively; P = .0136). Neurological complications were 9.5 times more common in the SBTKA group (1.58% vs 0.17%, respectively; P = .0024). All other complications were comparable in the 2 groups. CONCLUSION: Risk screening and preoperative optimization reduce mortality and overall complication rates in SBTKA patients; however, overall procedure-related complications, specifically cardiovascular and neurological, remain significantly high in SBTKA patients, for which a guarded approach is recommended.

Use of a mandibular torus for autogenous grafting: a case report.

Conventional treatment for the management of 2- to 3-walled intrabony defects is the use of allograft or alloplasts. Autogenous grafts are the gold standard because of their osteogenic potential. Mandibular tori are an ideal site for harvesting bone because their excision causes no structural, esthetic, or functional compromise to the patient. This case report describes the use of an autogenous graft obtained from a mandibular torus. The graft was utilized, in particulate form, to fill an intrabony defect at the mandibular right central incisor. The mandibular torus provided sufficient graft material and eliminated the need for a second surgical site. A follow-up at 1 year revealed reduction in clinical attachment loss and complete resolution of tooth mobility.

PMA and crystal-induced neutrophil extracellular trap formation involves RIPK1-RIPK3-MLKL signaling.

Neutrophil extracellular trap (NET) formation contributes to gout, autoimmune vasculitis, thrombosis, and atherosclerosis. The outside-in signaling pathway triggering NET formation is unknown. Here, we show that the receptor-interacting protein kinase (RIPK)-1-stabilizers necrostatin-1 or necrostatin-1s and the mixed lineage kinase domain-like (MLKL)-inhibitor necrosulfonamide prevent monosodium urate (MSU) crystal- or PMA-induced NET formation in human and mouse neutrophils. These compounds do not affect PMA- or urate crystal-induced production of ROS. Moreover, neutrophils of chronic granulomatous disease patients are shown to lack PMA-induced MLKL phosphorylation. Genetic deficiency of RIPK3 in mice prevents MSU crystal-induced NET formation in vitro and in vivo. Thus, neutrophil death and NET formation may involve the signaling pathway defining necroptosis downstream of ROS production. These data imply that RIPK1, RIPK3, and MLKL could represent molecular targets in gout or other crystallopathies.